• Journal of Oral Medicine and Pain
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• v.25 no.1
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• pp.29-39
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• 2000

Lysyl Oxidase from fetal bovine aorta was purified to homogenity using extraction, Sephacryl S200HR chromatography, Hydropore AX ion-exchange high performance liquid column chromatography, Cibacron blue affinity chromatography, and Sephacryl S-300 HR chromatography in the presence of protease inhibitor. The purified enzyme was active toward lathyritic collagen as well as elastin and was sensitive to aminonitriles such as BAPN. Upon Sephacryl S-300 HR chromatography, the enzyme was eluted as a peak with a $K_{av}$ value of 0.45 (65% of $V_t$ ) and it eluted from high performance liquid ion-exchange column (Hydropore |AX) at single position (ionic strength, I = 0.1~0.15). Once purified, it showed one band upon SDS-PAGE. It migrated to a band the mobility of which corresponded to a Mr of 33,500 upon reduction while it migrated to a 24,500 Mr position under the non-reducing condition. In constrast to other reports, it is concluded that fetal bovine aorta contains only one type of lysy oxidase.

• CELLMED
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• v.4 no.2
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• pp.11.1-11.6
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• 2014

Cinnamomum zeylanicum (C. zeylanicum) is a tropical evergreen tree of Lauraceae family. It is one of the oldest culinary spices known and used traditionally in many cultures for centuries. In addition to its culinary uses, cinnamon also possesses as a folk remedy of many health disease condition including analgesic, antiseptic, antispasmodic, aphrodisiac, astringent, carminative, haemostatic, insecticidal, and parasiticide and memory enhancing property. This study was aimed to assess the acetylcholinesterase and butyrylcholinesterase inhibitory activity of standardized methanol extract of the C. zeylanicum. Gas chromatography - mass spectrometry (GC-MS) and high performance liquid chromatography (HPLC) analysis were done to identify the presence of eugenol as chemical component and support the neuroprotective activity in the extract. Anticholinesterase inhibitory activity of crude methanol extract of C. zeylanicum leaves and cinnamon oil were evaluated by 96-well microtiter plate assay and thin layer chromatography bioassay detection methods. This study revealed that cinnamon oil ($IC_{50}:45.88{\pm}1.94{\mu}g/ml$) has better anticholinesterase activity than methanol extract ($IC_{50}:77.78{\pm}0.03{\mu}g/ml$). In HPLC analysis, retention time of eugenol in cinnamon oil was found to be 15.81 min which was comparable with the retention time (15.99 min) of the reference standard, eugenol. Seven chemical compounds were identified by GC-MS analysis, in which eugenol as an important phytoconstituents. Thus the phytochemicals from C. zeylanicum methanol leaves extract could be developed as potential source of anticholinesterase activity, with particular benefit in the symptomatic treatment of Alzheimer's disease.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.6 no.2
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• pp.202-208
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• 1996

The purposes of this study are the identification and determination of metabolites in the isolated rat liver perfusate of carbon disulfide by two-dimentional thin-layer chromatography and high performance liquid chromatography for understanding the metabolism of carbon disulfide. 2-Thio-thiazolidine-4-carboxylic acid(TTCA) was synthesized by the reaction of carbon disulfide and cysteine, and confirmed by two-dimentional thin-layer chromatography, high performance liquid chromatography, UV spectroscopy, and IR spectroscopy. The absorbance of UV detector for the simultaneous determination of TTCA and thiocarbamide was 254 nm although their maximum spectra were 273 nm and 237 nm, respectively. Two kinds of the developing solvent in the two-dimentional thin-layer chromatography were 2-butanol : 80% HCOOH : $H_2O$ (7 : 2 : 1) as the first developing solvent and 2-propanol : $H_2O$ (4 : 1) as the second developing solvent. After perfusion of carbon disulfide ($8274.23{\mu}mol$), the amount of TTCA and thiocabamide of the perfusate(100 ul) were $12.02-16.4{\mu}mol$ and $5.25-8.15{\mu}g$, respectively. The mean amount of them were $14.08{\mu}mol$ and $6.41{\mu}mol$ respectively, and the former was 2.20 times greater than the latter. For conforming the mechanism of formations of TTCA and thiocarbamide in vivo, we have to clarify whether the reactions between carbon disulfide and ammonia, ammonium salts, amides, cysteine, cystine, or proteins will be formed in vitro.

• Macromolecular Research
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• v.14 no.3
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• pp.383-386
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• 2006

High temperature size exclusion chromatography (SEC) has been used widely for the characterization of crystalline polymers, for which high temperature operation above the polymer melting temperature is required to dissolve the polymers. However, this high temperature operation has many advantages in SEC separation in addition to merely increasing polymer solubility. At high temperature the eluent viscosity decreases, which in turn decreases the column backpressure and increases the diffusivity of the analytes. Therefore, many reports on the high temperature operation of high performance liquid chromatography (HPLC) have focused on shortening the analysis time and enhancing the resolution. However, the application of high temperature SEC analysis to exploit the merits of high temperature operation is scarce. In this article, therefore, we report on a new apparatus design for high temperature SEC.

• Natural Product Sciences
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• v.21 no.1
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• pp.20-24
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• 2015

Cyanidin-3-glucoside (C3G) and cyanidin-3-rutinoside (C3R) were isolated by high-performance countercurrent chromatography (HPCCC) using a two-phase solvent system composed of tert-butyl methyl ether/n-butanol/acetonitrile/water/trifluoroacetic acid (1 : 3 : 1 : 5 : 0.01, v/v) to give pure C3G (34.1 mg) and C3R (14.3 mg) from 1.5 g crude mulberry fruit extract. Using the pure C3G and C3R, a reliable high-performance liquid chromatography (HPLC) method was developed and validated to determine the C3G and C3R contents in mulberry fruit. C3G and C3R were separated simultaneously using an Eclipse XDB-C18 column ($4.6{\times}250mm$ I.D., $5{\mu}m$) coupled with a photodiode array detector (PDA). The gradient elution of the mobile phase consisting of acetonitrile (0.5% formic acid) and water (0.5% formic acid) was applied (1.0 mL/min), and the detection wavelength was 520 nm. The calibration curves of C3G and C3R showed good linearity (both with $r^2=0.9996$) in the concentration range $15.625-500{\mu}g/mL$, and the relative standard deviations (RSD%) of intra- and inter-day variability were in the ranges 2.1 - 8.2% and 4.1 - 17.1%, respectively. The accuracies were ranged 96.5 - 102.6% for C3G and C3R, respectively. The developed HPLC method was used to determine the contents of C3G and C3R in newly harvested mulberry from eight different provinces of Korea.

• KSBB Journal
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• v.27 no.4
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• pp.199-206
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• 2012

Chiral crown ethers are synthetic macrocyclic polyethers that bind protonated chiral primary amines with high selectivity and affinity. They have been widely used to separate or distinguish the enantiomers of chiral compounds containing a primary amino moiety by high-performance liquid chromatography, capillary electrophoresis, and NMR spectroscopy. In this paper, two important chiral crown ethers including chiral binaphthyl unit and (18-crown-6)-2,3,11,12-tetracarboxylic acid as chiral selectors are focused. And several chiral resolution techniques and their applications in chirotechnology using these chiral crown ethers with related chiral recognition mechanism studies are reviewed. Especially, it was shown that the commercially available HPLC columns based on (18-crown-6)-2,3,11,12-tetracarboxylic acid have been developed and successfully applied for the resolution of various primary amino compounds including amino acids.

• YAKHAK HOEJI
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• v.36 no.5
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• pp.506-511
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• 1992

The Prostaglandins were derivatized rapidly with monodansyl cadaverine as a fluorophore in mild conditions. The carboxylic moiety of prostaglandins was activated with diethyl phosphorocyanidate and successively coupled with fluorophore in dimethylformamide at room temperature. The labeling yield was reached about 95% at 15 min using arachidic acid $(C_{20:0})$ as a test sample. This derivative showed constant fluorescent intensity at $4^{\circ}C$ for 180 days. The derivatives of prostaglandins were shown high solvent selectivity with tetrahydrofuran in reversed-phase column. therefore, these derivatives could be successfully separated on YMC pack A-212(S-5 120A C8) column in tetrahydrofuran-based eluents. The detection limits of these derivatives was ca. 500 fmol and determination limits was ca. 5 pmol as injected amount in fluorescent detection $({\lambda}ex.\;340\;nm,\;{\lambda}em.\;520\;nm)$. In this method, the ranges of recovery and coefficient of variation were $93.6{\sim}102.7%$ and $4.3{\sim}5.8%$, respectively.

• Food Science and Biotechnology
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• v.17 no.6
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• pp.1299-1304
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• 2008

The potential antioxidant activities of different fractions from a 70% methanolic (MeOH) extract of Sonchus oleraceus were assayed in vitro. All of the fractions exception of n-hexane showed a strong antioxidant activity, especially the ethyl acetate (EtOAc) fraction, which showed the highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity ($IC_{50}=19.25{\mu}g/mL$). The results of hydroxyl radical scavenging activity and a reducing power assay showed concentration dependence, the EtOAc fraction demonstrating a better result than the other fractions at the same concentration in the studies. Additionally, the fractions' total phenolic (TP) contents was measured, phenolic compounds such as tannic acid, p-coumatric acid, quercetin, epicathchin, and kaempferol being detected by high performance liquid chromatography (HPLC). Meanwhile, a regression analysis revealed a moderate-to-high correlation coefficient between the antiradical activity and the TP contents, suggesting that fractions obtained from the 70% MeOH extract of S. oleraceus are of potential use as sources of antioxidant material.

• Archives of Pharmacal Research
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• v.22 no.3
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• pp.294-299
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• 1999

Precolumn orthophthaldehyde (OPA) labeling method of sphingoid bases, sphingosine and sphinganine, was investigated to obtain high fluorescent detectability. In order to improve the fluorescent yield, we investigated the optimal solubility of sphingoid bases for five pre-incubation solvents by incorporating the heating procedure before OPA derivatization. The pre-incubation in ethanol prominently increased the fluorescent peak height of OPA derivative for each sphingoid bases in high performance liquid chromatography. About tenfold increase of detectability was archived by pre-incubating lipid extracts pellets in ethanol at $60^{\circ}C$ for 30 min. Optimal derivatization was performed in 30 min at ambient temperature and the fluorescent intensity of OPA derivative was stable for two weeks at $4^{\circ}C$. The detection limit of sphingosine was 0.1 pmol as injected amount. This method was applied to the determination of cellular sphingosine and sphinganine in various human lung cancer cells. This OPA procedure was prospective to be useful for quantitating the amount of sphingoid bases in other cancer cells.

• Food Science of Animal Resources
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• v.41 no.4
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• pp.731-738
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• 2021

Herein, a novel analytical method using a high-performance liquid chromatography-fluorescence detector (HPLC/FLD) is developed for rapidly measuring an L-carnitine ester derivative in infant powdered milk. In this study, solid-phase extraction cartridges filled with derivatized methanol and distilled water were used to effectively separate L-carnitine. Protein precipitation pretreatment was carried out to remove the protein and recover the analyte extract with a high recovery (97.16%-106.56%), following which carnitine in the formula was derivatized to its ester form. Precolumn derivation with 1-aminoanthracene (1AA) was carried out in a phosphate buffer using 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) as the catalyst. Method validation was performed following the AOAC guidelines. The calibration curves were linear in the L-carnitine concentration range of 0.1-2.5 mg/L. The lower limit of quantitation and limit of detection of L-carnitine were 0.076 and 0.024 mg/L, respectively. The intra- and interday precision and recovery results were within the allowable limits. The results showed that our method helped reduce the sample preparation time. It also afforded higher resolution and better reproducibility than those obtained by traditional methods. Our method is suitable for detecting the quantity of L-carnitine in infant powdered milk containing a large amount of protein or starch.